Molecular genetic analysis of hindlimb development in zebrafish

尽管人类先天肢体形成的复杂性和相对丰富性，但斑马鱼腹鳍/后肢形成的潜在和特有的分子机制在很大程度上还没有被探索。我们建议使用不同的遗传方法来揭示斑马鱼腹鳍/后肢形成的分子调控。为此，(I)我们将克隆和检测一个新的突变体，该突变体在腹鳍及相关内骨骼的形成中存在严重缺陷，(Ii)我们将测试RA信号对于腹鳍起始是必需的假设，并将研究与腹鳍内骨骼图案的意外相关，以及(Iii)我们将开发转基因工具，使我们能够以特定组织的方式，在任何给定的时间，操纵RA信号，并最终操纵任何信号途径。总之，我们的工作应该为脊椎动物后肢发育的调控提供新的和重要的见解。我们的发现将有助于更好地理解导致四肢先天性畸形的潜在机制。

Despite the complexity of human congenital limb formations and their relative abundance, the molecular mechanisms underlying and specific to pelvic fin/hindlimb formation in the zebrafish have largely been unexplored. We propose to unravel the molecular regulation of zebrafish pelvic fin/hindlimb formation using various genetic approaches. To this end, (I) we will clone and examine a novel mutant with severe defects in the formation of pelvic fins and associated endoskeleton, (II) we will test the hypothesis that RA signaling is required for pelvic fin initiation and will examine an unexpected involvement in patterning the pelvic fin endoskeleton, and (III) we will develop transgenic tools that allow us to manipulate RA signaling, and ultimately any signaling pathway, in developing pelvic fins in a tissue-specific manner and at any given time. Together, our work should provide new and significant insights into the regulation of vertebrate hindlimb development. Our findings should be relevant for a better understanding of the underlying mechanisms leading to congenital deformities of tetrapod limbs.